Data confirm that, under our experimental conditions, inhibition of MPO by 4-ABAH inhibits the formation of NETs. Therefore the product of MPO, HOCl was supplemented directly to neutrophils and was observed to elicit NET release PI3K inhibitor (Fig. 4a,b). This effect was found to be specific to the product of MPO as another chlorine-based acid (hydrochloric acid) evoked no detectable NET release (Fig. 4c). To confirm the physiological relevance of our hypothesis, we then exposed neutrophils obtained from patients with CGD to HOCl
to ascertain whether NET release could be evoked, despite the absence of a functional NADPH oxidase system (confirmed by chemiluminescent assay, data not shown). Neutrophils from CGD patients did not release NETs when stimulated with PMA, but were able to release NETs upon exposure to exogenous HOCl (Fig. 4d). Taurine is found abundantly within the cytoplasm of neutrophils (at ∼50 mM ) and is known to neutralize HOCl by forming taurine chloramine and essentially removing H2O2 and HOCl to promote cell survival . Indeed, taurine chloramine activates
Nrf2 and a battery of downstream cytoprotective anti-oxidant enzymes (including haem-oxygenase-1; glutathione-transferase; peroxiredoxin; thioredoxin), thus promoting cell survival . Therefore the role of taurine was examined by its addition prior to stimulation of NET release using both PMA (to stimulate endogenous HOCl generation) and also following direct addition of HOCl (0·75 mM). Ulixertinib in vitro Taurine treatment reduced NET release significantly in response to almost PMA at 100 mM and in response to HOCl at only 10 mM (Fig. 4e). This difference is likely to be due to both taurine and HOCl being added exogenously, and therefore the HOCl was likely to have been neutralized prior to entering the cell, unlike PMA which stimulates HOCl generation by direct intracellular activation of PKC. Direct neutrophil exposure to 0·75 mM HOCl resulted in the release of NET structures between 30 and 70 min (Fig. 5a), whereas stimulation with PBS did not
result in release of nuclear DNA (Fig. 5b) and treatment with 1% Triton X-100 killed neutrophils almost instantly to release non-NET DNA (Fig. 5c). The recent discovery of NET release  led to a plethora of studies describing their potential physiological role as a vitally important anti-microbial strategy in humans. However, their apparent complete dependence upon ROS activity suggests that the physiological heterogeneity surrounding ROS generation probably also pertains to NET release. Thus neutrophil hyperactivity and hyper-reactivity  with respect to ROS release may lead to disproportionate, physiologically discordant and/or displaced NET release with potentially pathogenic sequelae, such as autoimmune disease [8–10].